Project Summary The studies and career development/training activities in this K99/R00 proposal are designed to equip the PI, Dr. Rachel Perry, with the technical and scientific expertise and the experience to become an independent investigator exploring the topic of tumor metabolism. To that end, Dr. Perry will develop and optimize in vivo, ex vivo, and in vitro magnetic resonance and mass spectrometry methods to model glycolytic and oxidative metabolism in mouse models of colon and hepatocellular cancer, as well as in tumor-infiltrating T cells. These studies are designed to allow the identification of the mechanism(s) by which hyperinsulinemia ? which has been identified as a strong contributor to colon cancer risk and progression ? may drive tumor growth. Mice with colon carcinoma tumors will then be treated with a novel therapeutic agent, a controlled-release mitochondrial protonophore (CRMP), to reverse hyperinsulinemia, and its effect on tumor progression and metabolic flux rates will be identified, correlating altered substrate oxidation rates and/or insulin signaling markers in tumors with tumor growth. We will then treat mice with non-alcoholic steatohepatitis (NASH)- associated hepatocellular carcinoma (HCC) with CRMP. Because we have recently shown that this agent reverses NASH fibrosis, these experiments will test the hypotheses that reversing NASH will slow tumor growth, and that CRMP may be an attractive therapeutic option to slow HCC progression. We will also assess the alterations in metabolic flux rates that may occur in livers of mice with HCC as compared to normal livers using in vivo and ex vivo NMR/mass spectrometry techniques, and the effect that CRMP has on those fluxes. Because certain cancers have been associated with insulin resistance/hyperinsulinemia, non-alcoholic fatty liver disease, and NASH, the current obesity epidemic demands efforts to understand the mechanism(s) by which these factors may contribute to cancer pathogenesis, and the proposed studies have clear translational relevance. The work described above will be carried out by Dr. Perry in the Department of Internal Medicine/Section of Endocrinology at the Yale University School of Medicine, under the supervision of her mentor, Dr. Gerald Shulman, co-mentor Dr. Susan Kaech, and collaborators Drs. Douglas Rothman and Michael Pollak. The studies herein are carefully designed to broaden Dr. Perry's arsenal of technical skills as well as hone her scientific reasoning and provide career development training to enable her to become an Assistant Professor at the end of the K99 phase (after year 2 of the K99/R00 award), and to apply for independent R01 funding at the end of the R00 phase (in year 5 of the K99/R00 award). These goals will be achieved through Dr. Perry's plans, described in this application, to perform research; to meet frequently with her mentors, collaborators, and other members of the Yale faculty with research interests or technical skills relevant to these studies; to complete coursework; and to attend scientific meetings.